摘要
为建立在生物反应器内微载体逐级放大培养PK-15细胞和增殖PCV2技术。采用分批式培养方法,研究了PK-15细胞在微载体胰酶消化放大悬浮培养及猪圆环病毒2型(PCV2)增殖工艺。结果表明,5 g/L的微载体和高糖DMEM下,采用4.0×105cells/m L的初始接种密度及培养至第2天进行换液操作工艺可以获得最佳的PK-15细胞生长效能。胰酶消化转移将PK-15细胞从1 L反应器放大至3 L反应器,微载体上细胞贴附均匀、生长旺盛,培养3 d细胞密度可达34.3×105cells/m L。采用感染复数为0.1的接毒比例,细胞接毒后在微载体上传至第3代收获毒液可获得最高的PCV2增殖效价107.25TCID50/m L。为PCV2疫苗的生物反应器规模化生产奠定了基础。
In order to develop a technique for scale-up of microcarrier suspension culture of PK-15 cells and propagation of PCV2 in bioreae- tor,batch-wise suspension culture of PK-15 cells on microcarriers （ cytodex 1 ） using scale-up of tripsin digest and propagation of porcine cir- covirus type 2 （PCV2） were investigated. The results showed that the optimal growth efficiency could be achieved by 4.0 × 10^5 cells per 1 mL inoculation on 5 g/L concentration, DMEM medium with high glucose and media replacement after 2-day culture. The highest PCV2 titer was achieved 10^7.25TCIDso/mL at the 3^rd passage of PK-15 cells infected with 0.1 multiple of infection （MOI） of PCV2 The optimal processes achieved in this study laid the experimental foundation for mass production of PCV2 vaccines in bioreactor scale.
出处
《西南农业学报》
CSCD
北大核心
2017年第2期474-480,共7页
Southwest China Journal of Agricultural Sciences
基金
江苏省农业自主创新项目[CX(14)5044]
农业部公益性行业(农业)科研专项经费项目(201303046)
江苏省农业支撑计划(BE2012370)
